Compositions for linking zinc finger modules
First Claim
1. A zinc finger protein comprising a plurality of non-naturally occurring zinc finger modules, wherein at least two of the non-naturally occurring zinc finger DNA-binding modules are joined by an amino acid linker sequence of 5 to 20 amino acid residues between the last residue of the N-terminal zinc finger module and the first residue of C-terminal zinc finger module, the amino acid linker comprising an N-terminal amino acid residue adjacent to the N-terminal zinc finger module, a C-terminal amino acid residue adjacent to the C-terminal zinc finger module, and amino acid residues internal to the N- and C-terminal amino acid linker residues, wherein(i) the N-terminal amino acid linker residue or the internal amino acid linker residues comprises at least one proline residue;
- (ii) the zinc finger protein binds to a target site comprising target subsites bound by each zinc finger module; and
(iii) there is a gap of 0, 1, 2 or 3 base pairs between the target subsites bound by the zinc finger DNA-binding modules joined by the amino acid linker.
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Abstract
Disclosed herein are compositions for linking DNA binding modules to allow for specific and selective binding to module subsites separated by 1 or more base pairs. Also described are methods of making and using compositions comprising these linkers.
71 Citations
15 Claims
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1. A zinc finger protein comprising a plurality of non-naturally occurring zinc finger modules, wherein at least two of the non-naturally occurring zinc finger DNA-binding modules are joined by an amino acid linker sequence of 5 to 20 amino acid residues between the last residue of the N-terminal zinc finger module and the first residue of C-terminal zinc finger module, the amino acid linker comprising an N-terminal amino acid residue adjacent to the N-terminal zinc finger module, a C-terminal amino acid residue adjacent to the C-terminal zinc finger module, and amino acid residues internal to the N- and C-terminal amino acid linker residues, wherein
(i) the N-terminal amino acid linker residue or the internal amino acid linker residues comprises at least one proline residue; -
(ii) the zinc finger protein binds to a target site comprising target subsites bound by each zinc finger module; and (iii) there is a gap of 0, 1, 2 or 3 base pairs between the target subsites bound by the zinc finger DNA-binding modules joined by the amino acid linker. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification